Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Kit for simultaneous detection of multiple food-borne pathogenic bacteria and its application

A technology for food-borne pathogenic bacteria and pathogenic bacteria, which is applied in the directions of microorganism-based methods, microbial determination/inspection, microorganisms, etc. Problems such as few types of sex tests

Active Publication Date: 2020-12-04
JINAN KAICHEN BIOTEC
View PDF10 Cites 1 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] In order to make up for the deficiencies of the prior art and solve the cumbersome detection steps of food-borne pathogens in the prior art
For the problems of long detection period, inability to detect difficult-to-cultivate bacteria, and few types of one-time detection, the present invention provides a kit for simultaneous detection of multiple food-borne pathogens and its production method and application

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Kit for simultaneous detection of multiple food-borne pathogenic bacteria and its application
  • Kit for simultaneous detection of multiple food-borne pathogenic bacteria and its application
  • Kit for simultaneous detection of multiple food-borne pathogenic bacteria and its application

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0087] Embodiment 1: the extraction of sample nucleic acid

[0088] According to the national standard or industry standard method, the microorganisms in the food samples are enriched and cultured, and the bacterial DNA is extracted from the enrichment solution using a commercial bacterial genome DNA extraction kit. A total of 54 nucleic acid samples were extracted, of which 38 were positive samples with known subtypes, 10 were negative controls, and 6 were irrelevant bacterial samples. Specimen information is as follows:

[0089]

[0090]

Embodiment 2

[0091] Example 2: Preparation of Gene Chips for Parallel Detection of Foodborne Pathogenic Bacteria

[0092] First add a section of 8-20bp poly T to the 5' end of the above 19 probes, and carry out amino modification at the same time, dilute the probe with deionized water, and mix it with spotting solution in equal volume, so that the final concentration is 75pmol / ul , make the system dot matrix by cartesian microarray on the surface of the aldehyde-modified glass slide, place it at 75% relative humidity, and fix it at room temperature for 48-72 hours; after taking it out, put the slide in 0.2% SDS and shake and wash for 2min. Take it out and fully dry it at room temperature, then place the slide in sodium borohydride solution and shake for 5 minutes to remove free aldehyde groups; wash with 0.2% SDS for 1 minute, repeat twice, and finally soak in pure water for 2 minutes and dry in the air; The gene chip for parallel detection of food-borne pathogens with 19 probes is complet...

Embodiment 3

[0093] Embodiment 3: PCR amplification and hybridization

[0094] Add the above-mentioned sample nucleic acid (one cartridge to detect one sample, and a blank control is also provided as needed. In order to test the repeatability of the reagent, each sample is repeated three times) from the sample addition hole into the cartridge, and the cartridge All reagents required for PCR amplification, hybridization detection, and cleaning are pre-installed, and the genetic chip for parallel detection of pathogenic bacteria prepared in Example 2 is also pre-placed in the hybridization detection groove at the bottom of the cartridge. Put the cartridge into the cartridge processor, write the reaction program from the system control software, run it, and the instrument will automatically perform PCR amplification and hybridization.

[0095] (1) Qiagen Multiplex PCR Kit, Cat No.206143 was used for PCR amplification, two rounds of PCR amplification were performed, and the configuration of th...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses a kit capable of simultaneously detecting various food-borne pathogenic bacteria and an application of the kit, and belongs to the technical field of detection of food-borne pathogenic bacteria. The kit comprises parallel detection gene chips of the food-borne pathogenic bacteria fixed 19 pathogenic bacterium specificity probes, specific primers and universal primers. According to the kit, 19 typical food pathogenic bacteria such as campylobacter coli, campylobacter jejuni and enterobacter sakazakii can be simultaneously, rapidly and sensitively detected through reaction once, and detection data combined multiple amplification and multiple detection which cannot be provided by a traditional method are provided. According to a detection method, the time from sample treatment to generation of detection results only needs about 1-2 hours, accuracy rate is 100%, the method is simple in operation step, short in detection time, high in specificity, high in sensitivity and good in repeatability, 19 pathogenes causing foodborne diseases can be accurately detected in a classification manner, and the food-borne pathogenic bacteria are rapidly screened.

Description

technical field [0001] The invention relates to the technical field of detection of food-borne pathogens, in particular to a kit for simultaneously detecting multiple food-borne pathogens and an application thereof. Background technique [0002] Foodborne diseases caused by foodborne pathogens have become one of the most concerned public health problems in the world today. WHO lists Escherichia coli O157, Salmonella, Shigella, and Listeria monocytogenes as four important food-borne pathogens in sequence, and lists them as mandatory inspection items for China's import and export products. In my country, Salmonella, Staphylococcus aureus, and Vibrio parahaemolyticus are the most common pathogenic bacteria in food. According to the national food poisoning incident report of the Ministry of Health in 2013, the number of microbial food poisoning is the largest, accounting for 60.4% of the total number of food poisoning incidents, mainly caused by Salmonella, Vibrio parahaemolyti...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Patents(China)
IPC IPC(8): C12Q1/689C12Q1/6837C12Q1/14C12Q1/10C12Q1/04C12R1/19C12R1/42C12R1/445C12R1/63C12R1/01
CPCC12Q1/6837C12Q1/689C12Q2565/501C12Q2565/519C12Q2531/113
Inventor 李艳艳靖相密邱盟轩张通尚小云
Owner JINAN KAICHEN BIOTEC
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com